75N95022Q00287_SOW.pdf

Statement of Work (SOW)

A. GENERAL INFORMATION

1. Title of the Project: Fibrosis markers in kidney disease associated with dementia in women versus men.

2. Introduction and background:

The National Institute on Aging, NIH has a need for a project entitled, “Fibrosis markers in kidney disease associated with dementia in women vs. men”, to analyze the association of fibrosis markers and markers of chronic kidney disease (CKD) with cognitive function in the CKD patients, and specifically in women versus men.

The prevalence of chronic kidney disease (CKD) and cognitive impairment increases with age. CKD is a significant and independent somatic risk factor in the development of cognitive impairment, which is mirrored in animal models. In end-stage CKD (stages 4-5), the prevalence of cognitive impairment has been estimated at 30-60%, twice the values observed in age- and sex-matched controls. CKD is greater in females vs.

males due to a higher risk of renal infections, especially during the pregnancy, which may result in the renal complications later in life. Hormone replacement therapy may be beneficial for CKD outcomes in women.

Cardiovascular diseases (CVD) including hypertension and increased central arterial stiffness (CAS) accompany CKD development across all stages of this disease. Pulse wave velocity (PWV), which is an independent predictor of the CKD progression, is the best non-invasive measurement of CAS. Patients with CKD have increased risk of thrombosis, bleeding and susceptibility to venous thromboembolism. These events may contribute to the development of cognitive impairment because cognitive impairment persists in 30% of the patients with cerebral venous thrombosis. Older women have slightly higher risk of venous thrombosis. The burden of hemostasis biomarkers is relevant to cognitive impairment. Platelet activation is negatively associated with PWV, suggesting that CAS plays an important role in thrombotic events. One of the factors responsible for the increased PWV is tissue fibrosis.

Multiple pro-fibrotic factors, such as Angiotensin II (AngII) and an endogenous Na/K- ATPase steroidal inhibitor, marinobufagenin (MBG), are implicated in the development of CKD via their effects on blood pressure and CAS as demonstrated in both clinical studies and animal models. MBG binds to Na/K-ATPase and activates pro-fibrotic Fli1-and TGFβ-dependent signaling. Platelet TGFβ1 is the major contributor to cardiac fibrosis and systolic dysfunction in murine pressure overload models. Men and women exhibit different profile in MBG level and its association with high dietary salt intake and cardiovascular parameters.

MBG and other pro-fibrotic factors may be implicated in the development of vascular dementia in CKD via stimulation of CAS, which drives pulse pressure increases and cerebral microvascular damage resulting in a reduction of blood supply to the brain hereby contributing to cognitive impairment. Elevated pulse pressure is associated with higher mortality in CVD and CKD independently of blood pressure. Moreover, pulse pressure is a cardiovascular risk factor in women >55 years. Thus, increase in PWV and pulse pressure is implicated in age-associated changes in the brain and potentiates the development of cognitive impairment in CVD and CKD.

MBG levels are elevated in hypertension, preeclampsia, CKD, and in the animal models of these diseases, conditions that expedite aging and premature death. MBG is a pro-hypertensive and pro-fibrotic factor, which participate in the development of CVD in CKD in humans and in the animal models of CKD. Although sex difference in MBG production has been studied in humans and in animals, the pathological sex-associated impact of MBG in CKD is not fully investigated.

The 17β-lactone ring of steroid MBG is critical for Na/K-ATPase binding and can be hydrolyzed and inactivated by a family of lactonase enzymes Paraoxonases (PONs).

Decreased PON enzymatic activity in CKD, which is associated with elevated MBG levels, can predict adverse cardiovascular outcomes in CKD. Thus, in addition to MBG immunoneutralization, augmentation of PON lactonase activity may be a novel counter-regulatory mechanism to reduce MBG mediated pro-fibrotic signaling and improve clinical outcomes such as cognitive impairment in CKD.

3. Objectives:

This requirement will utilize datasets and samples from an ongoing Bench to Bedside (BtB) NIH clinical study on CKD that has been performed at Marshall University under the direction of Dr. Sodhi. In an endeavor to expand the understanding of the effect of renal disease on cognitive function, this project will measure the markers of arterial stiffening and the markers of neurodegeneration in the existing plasma samples. Renal disease may affect cardiovascular system and thus reduce the delivery of blood to brain. The investigation into age and sex of the patients is also needed, because older women with renal disease may be at higher risk of cognitive impairment. Finally, the analysis of therapeutical treatments of the subjects may provide additional information about the benefit of certain therapeutical approaches, which can be revealed with the suggested multi-layered study.

4. Type of Contract: Firm-Fixed-Price

B. CONTRACTOR REQUIREMENTS

In the performance of the tasks specified, the Contractor shall be responsible for properly protecting all information used, gathered, or developed in result of the present SOW. According to Government rules, the Contractor shall establish and implement appropriate administrative, technical, and physical safeguards to ensure the security and confidentiality of sensitive data. Including personal protected identification.

Requirements to are as follows:

Tasks 1 and 2: To perform a cross-sectional analysis of clinical study datasets with a consideration of sex as a biological variable in age- and education matched the existing databases for cardiovascular parameters and mini-mental status examination from 60 female and 60 male CKD patients (stage 4), and 60 female and 60 female healthy subjects will be obtained. The Contractor shall provide the data (values) for all measurements, described below, in electronic forms and reports. Statistical analyses shall be prepared for publications.

1. The Contractor shall study the association of plasma markers, which accompany the development of CKD and CVD, including MBG, ANGII, aldosterone, BNP, collagen turnover and thrombosis/stenosis markers in the CKD patients vs. healthy controls.

Clinical parameters, including blood pressure, echocardiography (ECHO), PWV, cognitive mini-mental status examination (MMSE), which included orientation, attention and calculation, recall, language and copy design shall be assessed. Contractor will take into consideration CKD datasets and other pharmacological therapies. These therapies will allow an estimation of the effect of different medications on cardiovascular parameters and cognitive function. The measurement of specific serum PON hydrolytic activity toward MBG and the measurements of fibrosis markers shall be performed on the existing samples.

2. The cross-sectional case-control study will utilize datasets from the patients, that attended Dialysis Clinic at Cabell Huntington Hospital. The datasets will be selected based on the inclusion criteria (for the male and female CKD cohort: age 45 years and older, CKD stage 4, GFR 15-29, and Creatinine > 1mg/dL; n=60 for each sex); for the control cohort: age 45 years and older, systolic blood pressure < 130 mmHg, diastolic blood pressure < 80mmHg, GFR > 90, and Creatinine < 1mg/dL; n=60) and the exclusion criteria (history of HIV, hepatitis, malignancies, admission and/or surgery for trauma, and head injury). The subjects in the dataset will be matched for educational attainment to assess the results confounded by difference in education.

3. Stored plasma samples from the Dialysis Clinic at Cabell Huntington Hospital (240 samples) will be analyzed for basal level of platelet activation as well as responses to agonist-stimulations will be examined using aggregometer and flow cytometry.

Samples (n=240) will be used to measure the plasma levels of uremic toxins (indoxyl sulfate, p-cresyl sulfate), fibrotic markers (MBG, ANGII, aldosterone, TGFβ, collagen-1, Fli-1), oxidative stress and inflammatory markers (malondialdehyde, myeloperoxidase, TNFα, IL-6), markers of cognitive function (BDNF, amyloid-beta and t-tau), and sex hormones (estrogen and progesterone; in females) using the commercially available immunoassays.

4. 2D Doppler and color flow imaging ECHO datasets will be analyzed for the evaluation of ejection fraction which will be calculated using the standard formula. Aortic PWV from SphygmoCor® XCEL PWA&PWV measurements will be analyzed to assess the CAS in the patients.

5. Datasets from patients that were subjected to MMSE will be clustered into 11 sub-scores, which include orientation, attention, calculation, recall, language, copy design, and will be analyzed for the presence of the score decrease in any specific cognitive test domain.

Table 1. Proposed data sets

Parameters to be analyzed: Comments:

Number of CKD patients (women, %): 120 (50%)

The data shall be chosen based on the inclusion and exclusion criteria; data will be analyzed using 2-way ANOVA and linear regression statistical analyses

Number of controls (women, %):

120 (50%)

The datasets will be based on the inclusion and exclusion criteria. The data will be analyzed using 2-way ANOVA and linear regression statistical analyses

Age (years): >45 The demographic information will be part of the database which will be analyzed using 2-way ANOVA and linear regression statistical analyses

Hand cuff Systolic and Diastolic Blood Pressure and Heart Rate

Database will be separated by contractor based on exclusion criteria; the data will be analyzed using 2-way ANOVA and linear regression statistical analyses

Echocardiography and pulse wave velocity

Will be a part of the database for cardiovascular parameters; the data will be analyzed using 2-way ANOVA and linear regression statistical analyses

Blood Glucose, Triglycerides, Urea Nitrogen, Creatinine, brain natriuretic peptide

Will be a part of the database for cardiovascular parameters; data will be analyzed using 2-way ANOVA and linear regression statistical analyses

Mini-mental test Simple cognitive test results will be a part of the database; the data will be analyzed using 2-way ANOVA and linear regression statistical analyses

Task 3: The Contractor shall analyze behavioral test data from the established animal models of CKD and the results of the analysis should report the effect of anti-fibrosis therapy in these animal models of CKD. Treatments from a monoclonal anti-MBG antibody shall be analyzed for fibrosis and neurodegeneration markers (angiotensin II, MBG, TGFb, MBG, collagen-1, Fli-1), oxidative stress and inflammatory markers (malondialdehyde, myeloperoxidase, TNFα, IL-6), markers of cognitive function (BDNF, amyloid-beta and t-tau) and behavioral tests results. The animal CKD models include partial nephrectomy CKD model. The Contractor shall provide the data (values), where analysis was performed as described below, in electronic forms and reports as specified.

1. The database from the animal model for CKD will be used to analyze the initial platelet adhesion and activation and time to occlusive thrombus formation; sex-associated difference should be statistically analyzed by 2-way ANOVA. Histological examination of vessel stiffness will be analyzed for the thrombotic changes in carotid arteries. The behavioral tests results, including Novel Object Recognition test, O-maze, open field and attention tests will be included to the database and analyzed. The analysis of plasma pro-fibrotic markers (ANGII, aldosterone, MBG, TGFβ, collagen-1, Fli-1), oxidative stress and inflammatory markers (malondialdehyde, myeloperoxidase, TNFα, IL-6), markers of cognitive function (BDNF, amyloid-beta and t-tau) and hormonal profile (estradiol) shall be assessed. The data will be further analyzed using 2-way ANOVA and linear regression statistical analyses

2. Serum provided by NIA/LCS from Sprague-Dawley rats (n=64; 4-mo old and n=64 12-mo old) will be used. The samples include serum from 8 groups: males and females, CKD and control, treatment, and sham. The plasma samples will be used in immunoassays to measure ANGII, aldosterone, MBG, TGFβ, collagen-1, Fli-1), oxidative stress and inflammatory markers (malondialdehyde, myeloperoxidase, TNFα, IL-6), markers of cognitive function (BDNF, amyloid-beta and t-tau).

C. GOVERNMENT RESPONSIBILITIES

The Government will provide data sets for the contractor’s review.

The Government will provide 128 rat plasma samples for analysis by the contractor.

The contractor shall provide written description of the results and of the methods, the data analysis, and experimental suggestions.

The contractor will NOT require access to the NIA Biomedical Research Center 251 Bayview Boulevard, Baltimore, MD 21224-6825

D. DELIVERY OR DELIVERABLES

The contractor shall report in LCS group meeting (the day will be announced) all results 90 days after award. The progress reports will be submitted monthly; the final report will be submitted at the end of the contract period.

1. Description of Tasks and Associated Deliverables:

a) Contractor shall report during meeting of the Laboratory of Cardiovascular Sciences (LCS) group all results.

b) Contractor shall meet virtually with LCS to review data and data sets.

c) Contractor shall provide the analyzed data set with tables and figures for publication.

2. Reporting Requirements

a) The contractor shall provide monthly progress reports within fifteen calendar days after the end of each month.

b) The contractor shall submit one final project report upon its completion electronically.

Deliverables will be reviewed and approved by the Contracting Officer’s Representative, Richard Telljohan.

E. TRAVEL

Travel is required for the purpose of presenting and discussing results at the following two scientific conferences:

1 American Society of Nephrology (ASN) Kidney Week (2022) 2 Alzheimer’s Association International Conference (2022)

F. CONTRACTOR EXPERIENCE REQUIREMENTS – KEY PERSONNEL

The key personnel shall have experience with chronic kidney disease research, and experience in clinical and basic science research with principal research focus in nephrology and cardiovascular diseases. The key personnel shall have a broad background in kidney and metabolic pathophysiology, with specific training and expertise in metabolic disturbances and chronic kidney disease. The key personnel shall be a member of scientific societies focused in medicine, nephrology, cardiovascular diseases, physiology, metabolic syndrome or pharmacology and must have a good-standing.

G. SECURITY REQUIREMENTS

Information security is applicable.

H. DATA RIGHTS

All data rights regarding the use, production, and disclosure of that data belong to the NIH and the Contractor. The Contractor has the publication rights to data generated under this contract with a co-authorship with NIH.